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A kind of preparation method of stable isotope labeled glucose

A stable isotope and glucose technology, applied in the fields of biology, chemical industry, and medicine, can solve the problems of high cost of C methanol, unfavorable production and utilization, and high equipment requirements, and achieve the effect of improving purity, high utilization rate, and simple process

Active Publication Date: 2021-07-16
SHANGHAI RES INST OF CHEM IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0014] Chinese patent No. ZL 201010144928.9 discloses a preparation 13 The method of C labeling glucose, using 13 C methanol as the isotope raw material, Pichia pastoris as the starting strain, synthetic 13 C-labeled trehalose, synthesized by acid hydrolysis 13 C-labeled glucose, this patented technology is mainly to produce glucose from purified trehalose through acid hydrolysis. The acid hydrolysis method not only requires high equipment and has a great impact on the environment, but also 13 C methanol has high cost and low utilization rate, so it is not conducive to industrial production and utilization

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] obtained by photosynthesis 13 C-labeled Chlorella pyrenoidosa is dried, take 10g of bacteria, add 200ml of ethanol solvent, use Soxhlet extraction until the solution is colorless, add water to wash and centrifuge the solution until the liquid is colorless, freeze the centrifuged bacteria at -80°C overnight , take out and melt at 40°C, add 10mg of liquefying enzyme, adjust the pH of the solution to 6.5, react at 90°C for 1.5h, then raise the temperature to 110°C to deactivate the enzyme, cool down, add 10mg of glucoamylase, adjust the pH of the solution to 4.5, 60°C React for 24 hours, centrifuge to obtain the supernatant, adjust the pH of the supernatant to 6.5, put the liquid on the D301 resin column, set the flow rate to 90ml / h, wash the resin column with water, collect the eluate, concentrate, decolorize with activated carbon, crystallize, and dry , to get 5.1g of 13 C-marked glucose product with a purity greater than 98%. In this embodiment, hydrochloric acid or s...

Embodiment 2

[0041] obtained by photosynthesis 13 C-labeled Chlorella pyrenoidosa is dried, take 10g of bacteria, add 200ml of acetone solvent, use Soxhlet extraction until the solution is colorless, add water to wash, and centrifuge the solution until the liquid is colorless, after centrifugation, the bacteria are ultrasonically treated for 30min, Add 200mg of liquefied enzyme, adjust the pH of the solution to 6.5, react at 90°C for 1.5h, then raise the temperature to 110°C to deactivate the enzyme, cool down, add 500mg of glucoamylase, adjust the pH of the solution to 4.5, react at 60°C for 24h, centrifuge to obtain the supernatant solution, adjust the pH of the supernatant to 6.5, put it on the D296 resin column, set the flow rate to 90ml / h, wash the resin column with water, collect the eluate, concentrate, decolorize with activated carbon, crystallize, and dry to obtain 4.9g of 13 C-marked glucose product with a purity greater than 98%. In this embodiment, hydrochloric acid or sodium ...

Embodiment 3

[0043] obtained by photosynthesis 13 C-labeled Chlorella pyrenoidosa is dried, take 10g of bacteria, add 200ml of ether solvent, use Soxhlet extraction until the solution is colorless, add water to wash, and centrifuge the solution until the liquid is colorless, and use a cell crusher to process the centrifuged bacteria. The cells were broken for 30 minutes, then 100 mg of glucoamylase was added, the pH of the solution was adjusted to 6.5, and the reaction was carried out at 90°C for 1.5 hours. , centrifuge to obtain the supernatant, adjust the pH of the supernatant to 6.5, put the liquid on the A30 resin column, set the flow rate to 90ml / h, wash the resin column with water, collect the eluate, concentrate, decolorize with activated carbon, crystallize, and dry to obtain 4.5 g's 13 C-marked glucose product with a purity greater than 98%. In this embodiment, hydrochloric acid or sodium hydroxide is used to adjust the pH of the liquid.

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Abstract

The invention relates to a method for preparing stable isotope-labeled glucose, which comprises the following steps: pretreatment of raw materials: 13 C-labeled Chlorella pyrenoidosa as isotopic raw material, using organic solvents for 13 C-labeled Chlorella pyrenoidosa is subjected to decolorization treatment, and the decolorization bacterium cell crushing treatment; starch liquefaction and saccharification reaction: after the pretreatment 13 C-labeled Chlorella pyrenoidosa undergoes a liquefaction reaction with a liquefaction enzyme, and then undergoes a saccharification reaction with a glucoamylase, separates the reaction liquid from solid to liquid, and collects the supernatant; 13 Separation of C-labeled glucose: The supernatant is subjected to resin decolorization treatment to obtain an eluate, which is concentrated, activated carbon decolorized and crystallized to obtain the 13 C labeled glucose. Compared with the prior art, the present invention has 13 The C-labeled glucose product has the advantages of high abundance and purity, mild process conditions, little impact on the environment, and high utilization rate of chlorella.

Description

technical field [0001] The invention relates to a method for preparing glucose, in particular to a method for preparing stable isotope-labeled glucose, which belongs to the technical fields of biology, medicine and chemical industry. Background technique [0002] Glucose (glucose) molecular formula C 6 h 12 o 6 , molecular weight 180, white crystal, easily soluble in water, sweet taste, melting point 146°C, structural formula as follows: [0003] CH 2 OH-CHOH-CHOH-CHOH-CHOH-CHO. [0004] It is the most widely distributed monosaccharide in nature and is widely used in medical infusions, biological raw materials, etc. [0005] 13 C-labeled glucose is a labeled compound of glucose, not only used for clinical diagnosis of diabetes, early cerebrovascular, disorders or dementia lesions; as a raw material, to produce labeled glucose derivatives and other labeled products; it can also be used as a scientific research tracer for comparative use widely. [0006] Ordinary gluco...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/14C12P19/02C07H1/06C07H3/02
CPCC07H1/06C07H3/02C12P19/02C12P19/14
Inventor 刘占峰徐建飞侯静华张亮任征
Owner SHANGHAI RES INST OF CHEM IND
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